Project Summary - Biophysics and Metabolic Imaging Core The Biophysics and Metabolic Imaging Core will provide the instrumentation, facilities and expertise required to meet the optical microscopy and imaging analysis needs of the three Projects detailed in this application, as well as those of any PSOC pilot or trans-network projects that develop as a result of this Center program. In particular, the Core will provide the imaging infrastructure and expertise required for imaging of the cellular metabolic state, tissue structural features, and the general visualization of the biological samples tested in this Center. The Biophysics and Metabolic Imaging Core is made up of four overall aims: Aim 1: We will provide instrumentation and analysis support for high resolution quantitative optical imaging-based measurements of cellular metabolic state using new genetically encoded metabolite-sensing indicators and label-free two-photon excited NADH/FAD ratio imaging. Aim 2: We will develop a set of novel hardware platforms that will introduce further improved methods for carrying out FRET-based metabolic sensor imaging and combined lifetime/polarization anisotropy NADH imaging. We will also enable high throughput, high content, and long time-course imaging of metabolism and cellular features on the single cell level in culture systems and tissue explants such as patient-derived xenographs (PDXs). Aim 3: We will construct and utilize a new type of super-resolution interference-based microscope with 10 nm axial resolution that is especially designed to probe the cell membrane/ glycocalyx ? substrate interactions that play a critical role in how cancer cells interact with their environment. Aim 4: We will provide the computational expertise and computer hardware for the image processing and analysis required to evaluate the image data acquired in Aims 1-3. The Biophysics and Metabolic Imaging Core is co-directed by two recognized experts in biological imaging and provides cutting-edge instrumentation and expertise that will be critical to the success the overall project. The Biophysics and Metabolic Imaging Core will closely interact with the three Projects and with the Tissue Microfabrication Core on applications of our existing technologies and the development of new technologies for task-specific metabolic and functional imaging, image-based biophysical modeling, and analytical image analysis.